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目的 探讨人未成熟卵母细胞适宜的培养体系及培养时间.方法 将卵母细胞-卵丘复合体(0CC)332枚和裸卵(DO)393枚随机置入TCM199和P1两种培养体系体外成熟.OCC体外培养48 h,观察其成熟情况.DO分别在24、30、48 h观察第1极体排出情况,计算体外成熟率.OCC和DO成熟后称为成熟的卵母细胞,成熟的卵母细胞行精子卵浆内注射技术(ICSI)受精,比较其在两种培养体系的受精率、卵裂率及囊胚形成率.结果 在TCM199和P1培养体系中,OCC的成熟率、成熟后成为成熟的卵母细胞的受精率和卵裂率的差异无统计学意义(P均>0.05),但成熟的卵母细胞的囊胚形成率为53.7%比37.8%(P<0.05).DO在TCM199和P1培养体系中体外培养24、30、48 h时成熟率的差异无统计学意义;由DO成熟后形成的成熟的卵母细胞在P1培养体系中的受精率、卵裂率和囊胚形成率高于其在TCM199培养体系中,但只有受精率差异具有统计学意义(73.5%比62.9%,P<0.05);DO在两种培养体系中体外培养48 h和30 h的成熟率均明显高于24 h(P<0.05),但48 h与30 h的成熟率相比无差异.结论 OCC适宜在TCM199培养体系的培养;DO适官在P1培养体系的培养.DO体外成熟形成成熟的卵母细胞的时间可以缩短至30 h.

Objective To investigate the suitable culture medium and culture time for human immature oocytes. Methods Three hundred and thirty-two oocyte-cumulus-complexes (OCCs) and 393 denuded oocytes (DOs) were randomly cultured in TCM199 culture system and PI culture system. The mature condition of OCCs was observed at time point of 48 h after culture in vitro. After DOs were cultured for 24 h, 30 h,48 h, expulsion of first polarbody was overviewed. Maturing rates(MR) of OCCs and DOs in vitro were calculated. Mature oocytes from OCCs and DOs were subjected to intracytoplasmic sperm injection (ICS1). Main outcomes were compared including fertilization rate (FR), eleaverage rate (CR) and blastulation rate (BR) between two culture systems. Results There was no statistical difference in MR of OCCs,CR and FR of mature oocytes from OCCs in TCM 199 and P1 culture system (all P > 0.05). The BR of mature oocytes from OCCs in TCMI99 culture system was higher than that of P1 culture system (53.7% vs 37.8%, P < 0. 05). For DOs when comparing TCM199 culture system with PI culture system,no significant difference was found in MR after cultured for 24 h,30 h and 48 h in vitro. The FR,CR and BR of mature oocytes from DOs in PI culture system were higher than those in TCM199 culture system,but only FR was significantly different (73.5% vs 62.9%, P <0.05). The MR of DOs in two culture systems was all significantly higher after culture in vitro for 30 h and 48 h than that for 24 h (P < 0.05), but MR was no difference at 48 h compared with 30 h. Conclusions OCCs are suitably cultured in TCM199 culture system whereas DOs in PI culture system. Culture time of human mature oocytes from DOs in vitro can be shortened to 30 h.